pHR-PGK-NO4-synNotch-Gal4VP64 vector (V014052)

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Price Information

Cat No.	Plasmid Name	Availability	Add to cart
V014052	pHR-PGK-NO4-synNotch-Gal4VP64	In stock, 1 week for quality controls	Buy one, get one free!

Two tubes of lyophilized plasmid will be delivered, each tube is about 5µg.

Basic Vector Information

Vector Name:: pHR-PGK-NO4-synNotch-Gal4VP64

Antibiotic Resistance:: Ampicillin

Length:: 11438 bp

Type:: Protein expression

Replication origin:: ori

Host:: Mammalian cells, Lentivirus

Promoter:: PGK

Growth Strain(s):: Stbl3

Growth Temperature:: 37℃

pHR-PGK-NO4-synNotch-Gal4VP64 vector Map

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Plasmid Protocol

1. Centrifuge at 5,000×g for 5 min.

2. Carefully open the tube and add 20 μl of sterile water to dissolve the DNA.

3. Close the tube and incubate for 10 minutes at room temperature.

4. Briefly vortex the tube and then do a quick spin to concentrate the liquid at the bottom. Speed is less than 5000×g.

5. Store the plasmid at -20 ℃.

6. The concentration of plasmid re-measurement sometimes differs from the nominal value, which may be due to the position of the lyophilized plasmid in the tube, the efficiency of the re-dissolution, the measurement bias, and adsorption on the wall of the tube, therefore, it is recommended to transform and extract the plasmid before using it

General Plasmid Transform Protocol

1. Take one 100μl of the competent cells and thaw it on ice for 10min, add 2μl of plasmid, then ice bath for 30min, then heat-shock it at 42℃ for 60s, do not stir, and then ice bath for 2min.

2. Add 900μl of LB liquid medium without antibiotics, and incubate at 37℃ for 45min (30℃ for 1-1.5 hours) with 180rpm shaking.

3. Centrifuge at 6000rpm for 5min, leave only 100μl of supernatant to resuspend the bacterial precipitate and spread it onto the target plasmid-resistant LB plate.

4. Invert the plate and incubate at 37℃ for 14h, or at 30℃ for 20h.

5. Pick a single colony into LB liquid medium, add the corresponding antibiotics, incubate at 220rpm for 14h, and extract the plasmid according to the experimental needs and the instructions of the plasmid extraction kit.

pHR-PGK-NO4-synNotch-Gal4VP64 vector Sequence

LOCUS       V014052                11438 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  Exported.
ACCESSION   V014052
VERSION     V014052
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 11438)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..11438
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        438..542
                     /label="AmpR promoter"
     CDS             543..1400
                     /label="AmpR"
                     /note="beta-lactamase"
     rep_origin      1574..2162
                     /label="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
     promoter        2408..2737
                     /label="SV40 promoter"
                     /note="SV40 enhancer and early promoter"
     intron          3871..3936
                     /label="small t intron"
                     /note="SV40 (simian virus 40) small t antigen intron"
     CDS             4066..4086
                     /label="SV40 NLS"
                     /note="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
     polyA_signal    4511..4645
                     /label="SV40 poly(A) signal"
                     /note="SV40 polyadenylation signal"
     LTR             4814..5447
                     /label="3' LTR"
                     /note="3' long terminal repeat (LTR) from HIV-1"
     misc_feature    5494..5619
                     /label="HIV-1 Psi"
                     /note="packaging signal of human immunodeficiency virus
                     type 1"
     misc_feature    6111..6344
                     /label="RRE"
                     /note="The Rev response element (RRE) of HIV-1 allows for
                     Rev-dependent mRNA export from the nucleus to the
                     cytoplasm."
     CDS             6529..6573
                     /label="gp41 peptide"
                     /note="antigenic peptide corresponding to amino acids 655
                     to 669 of the HIV envelope protein gp41 (Lutje Hulsik et
                     al., 2013)"
     CDS             6722..6763
                     /note="Protein Tat from Human immunodeficiency virus type 1
                     group M subtype B (isolate WMJ22). Accession#: P12509"
                     /label="Protein Tat"
     misc_feature    6856..6973
                     /label="cPPT/CTS"
                     /note="central polypurine tract and central termination
                     sequence of HIV-1"
     promoter        7033..7532
                     /label="PGK promoter"
                     /note="mouse phosphoglycerate kinase 1 promoter"
     CDS             9368..9808
                     /label="GAL4 DNA binding domain"
                     /note="DNA binding domain of the GAL4 transcriptional
                     activator"
     CDS             9845..9994
                     /label="VP64"
                     /note="tetrameric repeat of the minimal activation domain
                     of herpes simplex virus VP16 (Beerli et al., 1998)"
     misc_feature    10113..10701
                     /label="WPRE"
                     /note="woodchuck hepatitis virus posttranscriptional
                     regulatory element"
     LTR             10811..11044
                     /label="3' LTR (Delta-U3)"
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     promoter        complement(11069..11087)
                     /label="SP6 promoter"
                     /note="promoter for bacteriophage SP6 RNA polymerase"